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Cholesterol Detection Kit (cell-based)

Sensitive, simple staining for cellular unesterified cholesterol.
Catalog #: K587
$205.00

Product Details

Cat # +Size K587-100
Size 100 assays
Detection Method Light and Fluorescence Microscopy (Ex/Em = 340-380/385-470 nm).
Applications Screen/study/characterize stimulators/inhibitors that affect cholesterol transport and localization in a variety of cells.
Features & Benefits • Simple, easy to perform, histochemical method of identification of unesterified cholesterol.
• Includes U-18666A, a cholesterol transport inhibitor.
Kit Components • Fixative Solution
• Assay Buffer
• Staining Dye
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Cholesterol is an essential structural component of animal cell membranes. It is required to maintain membrane structural integrity and fluidity. Cholesterol also serves as a precursor for the biosynthesis of several hormones, bile acid and vitamin D. Plasma membrane contains majority of the cellular cholesterol (80-90%) whereas little cholesterol resides in endoplasmic reticulum and in mitochondrial membrane. Transport of intracellular cholesterol within the cells to different compartments is through vesicular and non-vesicular pathways. Defects in these transport processes can alter cellular cholesterol metabolism resulting in pathological conditions. Filipin III is widely used as a probe for sterol localization in membranes. Interaction with cholesterol alters Filipin absorption and fluorescence spectra. This assay provides a simple, easy to perform, histochemical method of identification of unesterified cholesterol.


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Can we use frozen samples with this assay?
Fresh samples are always preferred over frozen samples. However, frozen samples can also be used, provided, they were frozen right after isolation, were not freeze thawed multiple time (for which we recommend aliquoting the samples before freezing) and have been frozen for relatively short periods.
Can we use a different wavelength than recommended for the final analysis?
It is always recommended to use the exact recommended wavelength for the most efficient results. However, most plate readers have flexibility in their band width of detection in increments of +/- 10 nm. Depending on this flexibility range, you can deviate from the recommended wavelengths within limits.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact yopur regional BioVision distributor.
What is the shelf life of this kit?
This kit is good for 12 months from the date of shipment in the unopened form when stored at the appropriate temperature and appropriate conditions. After opening and reconstitution, some of the components in this kit are good for 2 months at -20°C. Please refer to the datasheet for storage information and shelf life of each of the components.
Why are my standard curve values lower than those shown on the datasheet?
There are multiple factors which influence the signals like the incubation times, room temperature, handling etc. In general, to increase the value of the standards, you can increase the incubation time. As long as the standard curve is linear, it should be fine to use, since all of your samples will also be measured under the same conditions on this curve.
How do I normalize my samples against protein concentration?
You can use a protein quantitation assay on the supernatants you get from cell/tissue lysates or with any other liquid sample in the assay buffer.
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
Should I make a standard curve for every expt I do, or is one curve/kit enough?
Yes, I would strongly recommend you to do the standards every time you do the expt. There is always a chance that something was done differently that day and we do not want any conditions to differ between standards and samples.
Can the kit work on bacteria or yeast cells?
The kit has been standardized for mammalian cells only